With global warming, high temperatures have become a major environmental stress that inhibits plant growth and development. Plants evolve several mechanisms to cope with heat stress accordingly. One of the important mechanisms is the Hsf (heat shock factor)–Hsp (heat shock protein) signaling pathway. Therefore, the plant transcription factor Hsf family plays important roles in response to heat stress. All Hsfs can be divided into three classes (A, B, and C). Usually, class-A Hsfs are transcriptional activators, while class-B Hsfs are transcriptional repressors. In potato, our previous work identified 27 Hsfs in the genome and analyzed HsfA3 and HsfA4C functions that promote potato heat resistance. However, the function of HsfB is still elusive. In this study, the unique B5 member StHsfB5 in potato was obtained, and its characterizations and functions were comprehensively analyzed. A quantitative real-time PCR (qRT-PCR) assay showed that StHsfB5 was highly expressed in root, and its expression was induced by heat treatment and different kinds of phytohormones. The subcellular localization of StHsfB5 was in the nucleus, which is consistent with the characterization of transcription factors. The transgenic lines overexpressing StHsfB5 showed higher heat resistance compared with that of the control nontransgenic lines and inhibitory lines. Experiments on the interaction between protein and DNA indicated that the StHsfB5 protein can directly bind to the promoters of target genes small Hsps (sHsp17.6, sHsp21, and sHsp22.7) and Hsp80, and then induce the expressions of these target genes. All these results showed that StHsfB5 may be a coactivator that promotes potato heat resistance ability by directly inducing the expression of its target genes sHsp17.6, sHsp21, sHsp22.7, and Hsp80.
Verticillium wilt is a major disease that severely affects eggplant production, and a new eggplant miRNA named miRm0002 identified through high-throughput sequencing was highly induced by Verticillium wilt infection. However, the miRm0002 function was still elusive. In this study, the sequence of the miRm0002 precursor was cloned and transgenic eggplants were constructed. In vivo inoculation test and in vitro fungistatic test showed that overexpressing miRm0002 lines were more resistant to Verticillium dahliae and inhibiting miRm0002 lines were more sensitive, compared to the wild-type (WT) control. Some physiological indicators were selected and the results showed that SOD, POD, and CAT activities were significantly increased in Verticillium wilt-infected overexpressing miRm0002 lines, indicating that the expression of miRm0002 activates the antioxidant system. QRT-PCR assay showed that the transcript expression of miRm0002 candidate target ARF8, a gene encoding auxin response factor was negatively related to miRm0002 in WT as well as transgenic eggplants. However, RLM-RACE mapping and degradome sequencing showed miRm0002 could not cleave the sequence of ARF8. Taken together, these data suggest that miRm0002 plays a positive role in the defense response of eggplant against Verticillium wilt.
Verticillium wilt is a vascular disease caused by Verticillium dahliae, which has a serious impact on the yield and quality of eggplant. MiRNAs regulates plant growth, development, environmental adaptation and other life activities by controlling the expression of target genes. Although miRNAs have been systematically studied by high-throughput sequencing in eggplant (Solanum melongena L.), their target genes are not identified systematically, which would provide the basis for exploring the mechanism of miRNA mediated gene regulation in response to Verticillium wilt of eggplant. In this study, degradome sequencing was performed using two libraries separately prepared from mock-inoculated and V. dahliae-inoculated seedlings of eggplants. A total of 122 target genes for 49 miRNAs families were identified. Of them, 62 target genes were significantly induced by V. dahliae, including 34 targets were found only in the inoculated seedlings library and 28 targets were shown to be differentially expressed between both two libraries. Further analysis showed that a conserved miRNA could target various genes ranging from one up to 15 or a target gene could be cleaved by 1–5 different miRNAs. QRT-PCR analysis on five miRNAs and their target genes confirmed the reliability of the degradome sequencing data. GO analysis revealed that target genes owned diverse biological functions and transcription regulation was the largest functional category. Analysis on KEGG showed that target genes were mainly involved in plant-pathogen interaction pathway and plant hormone signal transduction pathway to resist disease. This result will provide basic information for further studying the regulatory mechanism of eggplant miRNAs in response to Verticillium wilt infection.
Verticillium wilt, a soilborne disease caused by Verticillium dahliae (V. dahliae), can severely affect the yields of Solanaceae crops. In a previous study, it was observed in Solanum torvum (S. torvum) that protein phosphatase 5 (PP5) was induced by V. dahliae infection. To elucidate the function of PP5 more clearly, this study cloned an StPP5 cDNA from S. torvum by PCR. The cDNA contained an ORF of 1458 bp long encoding a putative protein of 485 amino acid residues with a predicted molecular mass of 54.63 kDa and a theoretical isoelectric point of 5.66. StPP5 protein contained a conserved PP domain and showed high similarity to other homologous members of the PP5 family from various plant species. The expression of StPP5 gene was upregulated after V. infection and reached its maximum value at 24 h in leaves. In order to clarify the role of StPP5, four transgenic tobacco plants expressing StPP5 were generated through Agrobacterium-mediated transformation and identified by PCR. In vitro culture assay showed that the growth of V. dahliae in PDA medium containing proteins extracted from the leaves of transgenic tobacco line P6 was inhibited, whose inhibition rate was 55.1%, higher than the non-transgenic control. These results indicated that StPP5 might be involved in plant defense against V. dahliae infection.